Highly efficient gene knockout by injection of TALEN mRNA into oocytes and host transfer in Xenopus leavis
| Project/Area Number |
25430089
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Hiroshima University |
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Principal Investigator |
Nakajima Keisuke 広島大学, 理学(系)研究科(研究院), 助教 (60260311)
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| Co-Investigator(Kenkyū-buntansha) |
YAOITA YOSHIO 広島大学, 理学研究科, 教授 (00166472)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | Host-transfer / TALEN / Genome editing / Targeted gene knockout / Xenopus / knockout / oocyte / host-transfer / DEADSouth / genome editing / knock out / host transfer / ゲノム編集 / ゼノパス / ノックアウト |
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| Outline of Final Research Achievements |
Zinc-finger nucleases, transcription activator-like effector nucleases (TALENs) and the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system are potentially powerful tools for producing tailor-made knockout animals. However, their mutagenic activity is not high enough to induce mutations at all loci of a target gene throughout an entire tadpole. In this study, we present a highly efficient method for introducing gene modifications at almost all target sequences in randomly selected embryos. In our method, the efficiency is further improved by injecting TALEN mRNAs fused to the 3’UTR of the Xenopus DEADSouth gene into oocytes, which are then transferred into a host female frog, where they are ovulated and fertilized. The addition of the 3’UTR of the DEADSouth gene promotes mRNA translation in the oocytes and increases the expression of TALEN proteins to near-maximal levels three hours post fertilization (hpf).
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Report
(4 results)
Research Products
(16 results)
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[Journal Article] Xenopus pax6 mutants affect eye development and other organ systems, and have phenotypic similarities to human aniridia patients2015
Author(s)
Takuya Nakayama, Marilyn Fisher, Keisuke Nakajima, Akinleye O. Odeleye, Keith B. Zimmerman, Margaret B. Fish, Yoshio Yaoita, Jena L. Chojnowski, James D. Lauderdale, Peter A. Netland, Robert M. Grainger
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Journal Title
Developmental Biology
Volume: 408
Issue: 2
Pages: 328-344
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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